Method of treating viral diseases

ABSTRACT

A medicament based on antibodies contains an activated form of monoclonal, polyclonal, or natural antibodies to interferon in low or ultra-low doses prepared by multiple consecutive dilutions and exposure to external factors, preferably in accordance with homeopathic technology. In order to obtain antibodies, human or heterologous interferon alpha, beta, or gamma, including recombinant interferon, is used; a mixture of various, mostly centimal, homeopathic dilutions being employed. A method of treating a pathologic syndrome, whose formation is affected by interferon, consists in the use of activated forms of antibodies to interferon alpha, beta, or gamma in low or ultra-low doses obtained by multiple consecutive dilutions and exposure to external factors.

FIELD OF THE INVENTION

The preset invention relates to the field of medicine and can be usedfor the treatment and prevention of acute and chronic infectiousdiseases, mostly of viral etiology, and for producing animmunomodulating effect.

BACKGROUND OF THE INVENTION

It is a well-known practice to use antibodies for the treatment ofpathologic syndromes (SU 1331598 A, A 61 K 39/00, 1984; SU 1730144 A1, C12 N 7/00, 1992).

Pharmaceutical preparations (sera, immunoglobulins) based on antibodiesemployed in therapeutic doses are also well known (see, for instance,Register of Pharmaceuticals in Russia, Encyclopedia of Pharmaceuticals(in Russian), Moscow, 2000, pp. 358-359).

However, the use of such preparations may be accompanied by undesiredside effects caused by the injection of substantial doses of foreignproteins into the body.

A method of the treatment of a pathologic syndrome (whose development isaffected by various forms of interferon) by the administration ofinterferon has been described (see, for instance, J. Jeger, Ed.,Clinical Immunology and Allergology (Russian translation), Meditsina,Moscow, 1990, Vol. 3, pp. 479-482).

DESCRIPTION OF THE INVENTION

The present invention is directed at obtaining an efficient and safepreparation with immunotropic activity and no tolerance formation byusing activated forms of antibodies.

The formulated objective is attained by making an agent containing anactivated (potentised) form of monoclonal, polyclonal, or naturalantibodies to interferon in low or ultra-low doses prepared by multipleconsecutive dilutions and by exposure to external impact, mostlyfollowing homeopathic technology.

After the activation (potentiation) of antibodies to interferon theultra-diluted solutions (or pharmaceutical carriers impregnated with theactivated antibodies) retain their biological (pharmaceutical) activitymanifested in modification of molecular-cellular and systemic effectsmediated by interferon or its receptors. Unlike the action ofphysiologic (non-activated) forms of antibodies to interferon there isno suppression of the activity of interferon.

The antibodies are obtained with the use of human or heterologousinterferon alpha, beta, or gamma, including recombinant interferon.

The agent prepared according to the present invention is a newimmunotropic pharmacologic preparation possessing pronounced specificpharmacologic activity, free of side effects, retention of therapeuticaction, environmental purity, and low manufacturing costs. In contrastto non-activated forms of antibodies used, among other forms, in smalldoses, the homeopathically activated (potentised) antibodies tointerferon feature an action that does not suppress the activity ofendogenous interferon; the activated antibodies act more oftensynergistically (unidirectionally) with interferon and enhance, amongother things, the induction of various forms of endogenous interferon.

It is preferred to use mixtures of different, mostly centimal,homeopathic dilutions.

The method of treating a pathologic syndrome, whose formation mechanismsare affected by interferon, consists in that activated forms ofultra-low doses of antibodies to interferon alpha, beta, or gammaprepared by multiple consecutive dilutions and exposure to externalimpact are used.

EMBODIMENTS OF THE INVENTION

The new medicament is mainly prepared in the following manner.

Recombinant interferon gamma expressed in the Escherichia coli cultureis purified by electrophoresis to at least 97% of the active substanceand used as an immunogen in hybridoma technology for obtainingmonoclonal antibodies. The latter are affinity purified bychromatography on Protein A.

The method of preparation of monoclonal antibodies is described, forinstance, in the book edited by G. Frimel, Ed., Immunological Methods,Moscow, Meditsina, 1987, pp. 9-33.

The isolated antibodies to the recombinant human interferon gamma aresubjected to consecutive multiple dilutions and to the impact of anexternal mechanical factor until ultra-low or low doses are obtained,for example, by the homeopathic potentisation procedure (see W. Schwabe,Homöopathisches Arzneibuch, Stuttgart, 1978). This procedure gives riseto a uniform decrease in the concentration through consecutive dilutionsof 1 volumetric part of the initial matter (antibodies) in 9 volumetricparts (for decimal dilution, D) or in 99 volumetric parts (for centimaldilution, C) of a neutral solvent with multiple vertical shaking of eachsolution; preferably, different containers for each subsequent dilutionare used. Finally, this procedure gives the required dose (potency).

The external treatment in the course of concentration reduction can alsobe executed by exposure to ultrasonic, electromagnetic, or otherphysical factors.

The resultant medicines are used mostly in the dosage forms anddilutions adopted in the homeopathic practice: as alcoholic and aqueoussolutions or as tablets (granules) prepared by impregnating the carriercontained in the dosage form by the potentised solution to saturation;also, the potentised solution can be added directly to a liquid dosageform.

EXAMPLE 1

In studies of the action of antibodies to interferon on the humoralimmune response we immunized mice by ram erythrocytes (thymus-dependentcorpuscular antigen). After that, the mice received per os a preparationcontaining homeopathically potentised polyclonal sheep antibodies tomurine interferon alpha (a-IFN) in a mixture of C12+C30 dilutions withina period of 5 days. On the first day of the 5-day course the mice alsoreceived single intraperitoneal injections of cyclophosphamide (½ of themaximal tolerable dose). Five days later an evaluation of the humoralimmune response parameters showed that the administration of potentisedantibodies to a-IFN favored the growth of the functional activity ofantibody-forming cells in the spleen and an increase in the titres ofhemagglutinins in the blood serum, which was particularly importantagainst the background of immunosuppression. Thus, the claimed agentstimulates the humoral immune response.

EXAMPLE 2

In studies of the action of activated forms of ultra-low doses ofantibodies to interferon on the manifestation of the delayed-typehypersensitivity reaction (DHR) of mice we immunized the animals bysubcutaneous injection of ram erythrocytes (RE); the challenging dose ofRE was injected into the cushion of the rear paw at the end of a 10days' course of peroral administration of a preparation containinghomeopathically potentised monoclonal antibodies (AB) to the murineinterferon beta (β-IFN) in a mixture of C12+C30+C200 dilutions in anamount of 0.2 ml.

Simultaneously, we injected saline into the contralateral paw in thesame volume. The intensity of the response was assessed 24 hours laterfrom the reaction index (RI), which was calculated individually for eachanimal from the formula:

RI(%)=(P _(o) −P _(c))/P _(c)×100

where P_(o) is the weight of the test paw, P_(c) is the weight of thecontrol paw.

Table 1 presents the experimental data.

TABLE 1 Effect of Potentised AB to β-IFN on the Intensity of the DHRTest groups Indices RE AB to β-IFN + RE DHR RI 28.00 ± 2.34 36.60 ± 2.00P_(cl) = 0.026

It follows that AB to β-IPN activate the function of T-effectors, whichis manifested in the enhancement of the delayed-type hypersensitivityreaction in response to sensitization by ram erythrocytes.

EXAMPLE 3

In studies of the action of activated forms of ultra-low doses ofantibodies to interferon on the phagocytal activity of neutrophils inthe peritoneal exudate we evaluated the phagocytal activity ofneutrophils 24 hours after the end of a 10 days' course of subcutaneousadministration of natural antibodies (AB) to interferon gamma (γ-IFN) ina C60 homeopathic dilution in an amount of 0.1 ml.

The phagocytal activity was evaluated from the capacity of these cellsto absorb daily culture of St. aureus, Strain 209 (the concentration ofthe suspension of the microorganisms was 100×10⁶/ml. We took account ofthe neutrophils that had had absorbed the microorganisms (the phagocytalindex, PI) and the average number of staphylococci absorbed by one cell(the phagocytal number, PN).

TABLE 2 Effect of AB to γ-IFN on the Phagocytal Activity of NeutrophilsTest groups Indices RE AB to γ-IFN PI, % 8.80 ± 1.36* 29.20 ± 3.20P_(cl) = 0.0004 PN 7.61 ± 1.24   8.78 ± 2.33

These results indicate that AB to γ-IFN stimulate phagocytosissubstantially as compared with the control (RE immunization) because ofan increase in the proportion of neutrophils capable of absorbingstaphylococci.

EXAMPLE 4

Patient K. (female), aged 62, had been suffering from repeated fits offever accompanied by eruptions on the chest and pain along theintercostal nerves. Diagnosis: ‘relapsing herpes zoster’. The generallyadopted therapy (antiviral agents, analgetics) did not bring stableresults. As a result of everyday intake of potentised monoclonalantibodies to the human interferon gamma (C 1000, 1 tablet, twice aday), the fever and pain syndrome' were arrested on the third day. Onthe seventh day of the treatment the eruptions virtually disappeared.Recommendation: a preventive intake of the remedy, 1 tablet every otherday. No relapses of herpes have been noted within a year.

EXAMPLE 5

Patient Sh. (male), aged 5, had been regularly examined because ofrelapsing infections of the upper respiratory tract (rhinolaryngotracheitis lasting 10 days every 2 months). The assessment of hisimmune status revealed the lowering of the count of CD4 lymphocytes,neutrophils, and interferon gamma in the peripheral blood. Prescription:potentised polyclonal (immune) antibodies to the recombinant humaninterferon gamma in a mixture of D24+C30+C200 dilutions, 1 tablet everyday. Within a period of four months the child had no relapses of acuterespiratory infection; his parents say that the child became more activeand regained weight. The second evaluation of the immune status 6 monthsafter the beginning of the treatment indicated the normalization of theindices of the cellular and humoral immunity.

EXAMPLE 6

Patient P. (male), aged 34, complained of rhinitis, pain in thenasopharinx, and subfebrile condition. Diagnosis: acute viralrespiratory infection. The intranasal administration of drops of apotentised C12 aqueous solution of monoclonal antibodies to recombinanthuman interferon gamma (3 times a day) normalized patient's conditionwithin 2 days. A new examination revealed no catarrhal phenomena.

EXAMPLE 7

Patient S. (male), aged 32, was hospitalized in the infectious unit onthe second day of the disease with the diagnosis “influenza with a gravecourse”. Later on, the diagnosis was confirmed virologically. Thecondition on entry was grave, hyperthermia (up to 41.6° C.), mentalconfusion, heavy breathing, dry cough. Auscultation revealed vesicularbreathing in the lungs. Prescription: monoclonal antibodies to the humaninterferon gamma in mixed C12+C30+C200 potences, 1 ml of an aqueoussolution subcutaneously every 2 hours. Within 6 hours the bodytemperature fell to 37.4° C., the general condition became satisfactory.During subsequent 3 days the patient received the preparation twice aday, parenterally (1 ml of an aqueous solution). On the fourth day ofthe therapy the condition was satisfactory, the body temperature was36.4° C.; the patient noted general weakness. The patient was dischargedin a satisfactory condition on the fifth day after hospitalization.

EXAMPLE 8

Patient U. (male), aged 27, appealed to the urologist with complaintsabout unpleasant sensations, itch, pain in the urethra, frequentmicturate urges, mucosal discharge of one-month duration. A detailedinquiry also revealed pain in the ankles and muco-purulent dischargefrom the eyes. A visual examination detected hyperemia, edema, andadhesion of the urethral lips. Laboratory immunofluorescence studies ofthe urethral discharge detected Chlamydia trachomatis. Prescription: amixture of polyclonal antibodies to the human interferons alpha andgamma in a C 1000 homeopathic dilution, 1 tablet every day over a periodof 2 weeks. On the fifth day of the treatment the patient noted apronounced improvement of the symptoms and the lessening of thedischarge from the urethra and eyes. At the end of therapy theimmunofluorescent analysis of the prostatic secretion for Chlamydiatrachomatis gave a negative result. Recommendation: administration ofthe preparation every other day over a period of two weeks. The secondexamination showed that the patient had no complaints; no signs of thefrontal urethritis were found.

1-5. (canceled)
 6. A method of treating a viral disease which comprisesadministering to a subject suffering the disease, at least onehomeopathically potentized form of and antibody to interferon.
 7. Themethod of claim 6, wherein said at least one homeopathically potentizedform of antibody is administered in the form of one or more homeopathicdilutions.
 8. The method of claim 7, wherein said one or more of thehomeopathic dilutions comprises one or more centesimal homeopathicdilutions.
 9. The method of claim 8, wherein said one or morehomeopathic dilutions comprises a mixture of C12, C30, and C200homeopathic dilutions.
 10. The method of claim 6, wherein said at leastone homeopathically potentized form of antibody is to alpha, beta orgamma interferon.
 11. The method of claim 6, wherein said at least onehomeopathically potentized form of antibody is selected from monoclonal,polyclonal, or natural antibodies.
 12. The method of claim 6, whereinsaid at least one homeopathically potentized form of and antibody isobtained by multiple consecutive dilutions and exposure to externalfactors.
 13. The method of claim 6 wherein the viral disease is upperrespiratory tract viral disease, viral influenza or herpes.
 14. Themethod of claim 13 wherein the virus is influenza.
 15. The method ofclaim 13 wherein the virus is herpes.